***** Final - August 3, 2012 *****



# Significant New Alternatives Policy Program

**Foam -**  **Blowing Sector Risk Screen on Substitutes for**  **CFC -**  **11 ,**  **HCFC -**  **141b , and HCFC**  **- 22**  **in Rigid Polyurethane and Polyisocyanurate Laminated Boardstock**  **; Rigid Polyurethane Appliance ,**  **Commercial**

**Refrigeration, Spray,**  **and Slabstock; Integral Skin Polyurethane**  **Substitute: T** 

**rans -**  **1 -**  **c hloro**  **- 3,3,3**  **- trifluoroprop**  **- 1**  **- ene (**  **SolsticeTM 1233zd(E) )**  **This risk screen does not**  **contain Clean Air Act (CAA) Confidential Business Information (CBI)**  **and, therefore, may be disclosed to the public.** 

### 1. **INTRODUCTION** Ozone - depleting substances (ODS) are being phased out of production in response to a series of diplomatic and legislative efforts that have taken place over the past two decades, including the Montreal Protocol and the Clean Air Act Amendments of 1990 (CAAA). The U.S. Environmental Protection Agency (EPA), as authorized by Section 612 of the CAAA, administers the Significant New Alternatives Policy (SNAP) Program, which identifies acceptable and unacceptable substitutes for ODS in specific end -uses based on assessment of their health and environmental impacts. EPA’s decision on the acceptability of a substitute is based on the findings of a screening assessment of potential human health and environmental risks posed by the substitute in specific applications. EPA has already screened a large number of substitutes in many end-uses within all of the major ODS -using sectors, including refrigeration and air conditioning, s olvent cleaning, foam blowing, aerosols, fire suppression, adhesives, coatings and inks, and sterilization. The results of these risk screens are presented in a series of Background Documents that are available in EPA's docket. The purpose of this risk screen is to supplement EPA’s Background Document on the foam blowing sector (EPA 1994) (hereinafter referred to as the Background Document). This risk screen evaluates the potential use of trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene ( also know as (E) 1 - c hloro - 3, 3,3 -trifluoropro - 1 - ene and hereinafter referred to as SolsticeTM 1233zd(E) ) as a substitute for CFC - 11 , HCFC - 141b , and HCFC - 22 in rigid polyurethane and polyisocyanurate laminated boardstock; rigid polyurethane appliance, commercial refrigeration, spray, and slabstock; and integral skin polyurethane. Table 1 presents information on the proposed substitute. **Table 1**  **. Composition of SolsticeTM 1233zd(E)**  **Constituent Chemical Formula**  **CAS Number Percent of Total**  **(by weight) a**  trans- 1 - c hloro - 3,3,3 - trifluoroprop - 1 - ene C 3 H 2 ClF 3 102687 - 65 - 0 >99% **Potential Impurities (maximum concentration)******* Final - August 3, 2012 *****

The potential risks associated with use of foam blowing agents have been examined at length in the Background Document. The reader is referred to this reference for a detailed discussion of the methodologies used to conduct this risk screen. This risk screen addresses flammability risk and assesses the impact of potential releases of SolsticeTM 1233zd(E) on the atmosphere and climate. O ccupational exposure was modeled to ensure that manufacture and application of the proposed substitute in the proposed foam blowing end -uses does not pose unacceptable risk to workers. Lastly, general population exposure modeling was performed to ensure that the prop osed substitute would not pose an unacceptable risk to the population at large. Section 2 of this report summarizes the results of the risk screen for the proposed substitute. The remainder of the report is organized into the following sections:

 Section 3: Atmospheric Assessment  Section 4: Flammability Assessment  Section 5: Potential Health Effects  Section 6: Occupational Exposure Assessment  Section 7 : General Population Assessment  Section 8 : Volatile Organic Compound Assessment  Section 9 : References 

### 2. **S UMMARY OF RESULTS**  SolsticeTM 1233zd(E) is recommended for SNAP approval as a foam -blowing agent in the following enduses: rigid polyurethane and polyisocyanurate laminated boardstock; rigid polyurethane appliance, commercial refrigeration, spray, and slabs tock; and integral skin polyurethane . EPA's risk screen indicates that the use of the proposed substitute will be less harmful to the atmosphere than the continued use of CFC - 11, HCFC - 141b , and HCFC - 22 . Given that appropriate personal protective equipment (PPE) ( OSHA Category C or higher) and engineering controls (e.g., exhaust ventilation, vapor- in - air detection systems) will be used during installation, maintenance, and disposal activities, no significant toxicity risks to workers or the general populatio n are expected . In addition, based on the characteristics of the proposed substitute and the proper precautions listed in the MSDS for SolsticeTM 1233zd(E), the proposed substitute is not expected to present a significant flammability concern during manufacture or end -use (see Section 4). 

### 3. **ATMOSPHERIC ASSESSMENT** This section presents an assessment of the potential risks to atmospheric integrity posed by the use of SolsticeTM 1233zd(E) in the foam blowing sector. The ozone depletion potential (ODP), global warming potential (GWP), and atmospheric lifetime (ALT) of the proposed substitute are presented in Table 2 . As compared to CFC - 11 and HCFC - 141b, and HCFC - 22 , SolsticeTM 1233zd(E) is substantially less harmful to the ozone layer, has less climate impact, and a shorter atmospheric lifetime. . Inaddition, SolsticeTM 1233zd(E) also has lower climate impact and a shorter atmospheric lifetime than those predicted for other substitutes examined in the Background Document. SolsticeTM 1233zd(E) has a non - zero ODP, but at 0 .00024, this value is very small when considering continued used of HCFC - 141b or HCFC - 22 . A recent report by Wang et. al. (2011) modeled the potential of SolsticeTM 1233zd(E) to affect the amount of ozone in the global atmosphere. This report found that the release of all SolsticeTM 1233zd(E) , assuming it were substituted for all compounds it might replace and that the entire amount used were released , would have a statistically zero impact on global atmospheric ozone (less than 0.01% change in total column ozone). Thus, EPA beli eves that use of SolsticeTM 1233zd(E) would result in substantially less harm to the climate and ozone layer than the continued use of ozone - depleting substances ( ODS ) .***** Final - August 3, 2012 *****

**Table 2**  **. Atmospheric Impacts of SolsticeTM 1233zd(E)**  **Compared to CF**  **C -**  **11 ,**  **HCFC -**  **141b , HCFC**  **** **22, and HFC -**  **134a Ref** **rigerant ODP**  **GWP ALT**  SolsticeTM 1233zd(E) 0.000 2 4 - 0.00034 a ,c 4.7 - 7 b , a 26 days a CFC - 11 1.0 d 4, 750 e 45 years e HCFC - 141b 0. 12 d 725 e 9.3 years e HCFC - 22 0.13 d 1,810 e 12 years e HFC - 134a 0 1,430 e 14 years e a Assuming emissions occur in the major regions of the world where blowing agents that HBA - 2 might replace are significantly used. Wang et. al. (undated). b Wang et. a l. (2011) c SolsticeTM 1233zd(E) SNAP Submission (Anonymous Submitter 2011b ) d WMO (2011) e IPCC 4 th Assessment Report (Forster et al. 2007) 

### 4. **FLAMMABILITY ASSESSMENT** SolsticeTM 1233zd(E) is not flammable at ambient temperatures and atmospheric pressure, but can ignite when mixed with air under pressure and exposed to strong ignition sources; also, containers of SolsticeTM 1233zd(E) may rupture when exposed to heat (Anonymous Submitter 2011b). SolsticeTM 1233zd(E) is not expected be used under pressure, but may be exposed to a potential source of ignition during roofing applications. During such applications, tar, which may be used to construct the roof, is applied at temperatures ranging from 330° F to 440° F. Due to tar’s flashpoint of ≥450°F, there is a risk of ignition under these conditions. In order to mitigate this risk, it is important to carefully monitor the temperature to prevent the tar from overheating or igniting. . In the c ase of roofing applications, containers of tar should be equipped with temperature control devices, and that personnel applying tar do so in stages . Foaming applications should be coordinated with tarring applications to ensure that risks are reduced. By keeping Solstice TM 1233zd(E) away from all sources of ignition and high heat and adhering to the other precautions listed in the proposed substitute’s MSDS, Solstice TM 1233zd(E) does not pose a significant flammability risk . In addition, SolsticeTM 1233zd(E ) is expected to be incorporated into a typical polyurethane or polyisocyanurate foam formulation consisting of up to 15% flame retardants and other materials such that the final concentration of SolsticeTM 1233zd(E) in the manufactured foam is generally expected to be between 10% and 25% of the entire formulation by weight, further reducing flammability risk. B y following the proper precautions specified by the proposed substitute’s MSDS, it is unlikely that SolsticeTM 1233zd(E) will pose a significant flammability risk. 

### 5. **POTENTIAL HEALTH EFFECTS** To assess potential health risks from exposure to this proposed substitute, the submitter developed an acceptable exposure limit (AEL) of 300 ppm 1 for SolsticeTM 1233zd(E) (Anonymous Submitter 201 2 a). The AEL represents the maximum 8 -hour time weighted average (TWA) at which a worker can be exposed regularly without adverse effects.

1 An AEL of 1,000 ppm was independently developed for SolsticeTM 1233zd(E) based on review of available toxicity studies (see Appendix A). However, as the AEL value of 300 ppm determined by the submitter is more conservative, for purposes of this risk screen, 300 ppm was used to evaluate the potential occupational expo sures.***** Final - August 3, 2012 *****

According to the MSDS, exposure to SolsticeTM 1233zd(E) may be harmful by ocular or dermal absorption, inhalation, or ingestion. Exposures of SolsticeTM 1233zd(E) to the eyes will cause serious eye irritation and may cause frostbite. In case of ocular exposure, the MSDS for SolsticeTM 1233zd(E) recommends that person(s) should immediately flush the eyes, including under the eyelids, with water for 15 minutes; should frostbite occur, the water should be lukewarm, not hot. Medical attention should be sought if irritation develops or persists. Exposures of SolsticeTM 1233zd(E) to the skin may cause irritation or frostbite. In the case of dermal exposure, the MSDS for SolsticeTM 1233zd(E) recommends that person(s) should immediately wash the affected area with water and remove all contaminated clothing; should frostbite occur, bathe ( do not rub) the affected area with lukewarm, no t hot, water. If water is not available, cover the affected area with a clean, soft cloth. Medical attention should be sought if irritation develops or persists. SolsticeTM 1233zd(E) vapors are heavier than air and cause suffocation by reducing oxygen available for breathing, causing asphyxiation in high concentrations. If person(s) are exposed to high concentrations, the person(s) will likely not realize that he/she is suffocating, but may experience central nervous system effects, such as drowsiness and dizziness. If SolsticeTM 1233zd(E) is inhaled, person(s) should be immediately removed and exposed to fresh air. The MSDS recommends that if breathing is difficult, person(s) should be given oxygen, provided a qualified operator is present, and medical attention be sought. SolsticeTM 1233zd(E) is not likely to be hazardous by ingestion; however, in case of ingestion, the MSDS recommends having the person(s) drink a cup of water, if fully conscious, and consulting a physician immediately . D o not induce vomi ting without medical advice. EPA’s review of the human health impacts of this proposed substitute is contained in the public docket for this decision. T he potential health effects of SolsticeTM 1233zd(E) are unlikely to occur when following good industrial hygiene practices and the PPE and engineering control (e.g., ventilation) recommendations outlined in the MSDS for SolsticeTM 1233zd(E) and Sections 6 and 7 of this risk screen. 

### 6. **OCCUPATIONAL EXPOSURE ASSESSMENT** This section presents estimates of potential occupational exposures to SolsticeTM 1233zd(E) , which is anticipated to occur during application of spray foam and manufacture of blown foam. To ensure manufacture and application of SolsticeTM 1233zd(E) for the proposed foam blowing end -uses do n ot present a risk to workers, occupational exposure modeling was performed for the proposed substitute. Limited data are available regarding employee exposures to blowing agents for the proposed foam types. The submitter, however, was able to provide monitoring data on potential exposures in typical settings for HFC - 245fa . Due to their similar physical properties (i.e., boiling point, vapor pressure, heat of vaporization, and molecular weight), EPA believes that using HFC - 245fa as a surrogate provides a reasonable estimate of potential air concentrations of SolsticeTM 1233zd(E) (see Table 3). **Table 3**  **. Boiling Point and Vapor Pressure for SolsticeTM 1233zd(E)**  **and HFC -**  **245fa Analog**

**Parameter SolsticeTM 1233zd(E)**  **Proposed Substitute**  **HFC -**  **245fa Analog** Boiling Point 66  F 62.4  F Vapor Pressure 16 PSIA (at 68  C) 18 PSIA (at 68  C) Heat of Vaporization 83.4 Btu/lb (at bp) 84.6 Btu/lb (at bp) Molecular Weight 130.5 134 P otential occupational exposures to SolsticeTM 1233zd(E) at foam manufacture and spray foam application were modeled b ased on the HFC - 245fa monitoring data, and found not to present a***** Final - August 3, 2012 *****

significant risk to workers, as described in the following sections. 

### 6.1. **Occupational Exposure at Manufacture** Using the surrogate HFC - 245fa monitoring data provided by the submitter on boardstock and bunstock, appliance foam and laminated boardstock manufacture , maximum potential exposures to SolsticeTM 1233zd(E) were modeled for employees involved in the manufacture of foam. Table 4 presents highest exposure concentrations modeled for each foam type , based on the HFC - 245fa surrogate data. **Table 4. Maximum Modeled Occupational Exposures to**  **SolsticeTM 1233zd(E) During Foam**  **Manufacture End**  **- Use**  **Maximum Modeled Instantaneous**  **Exposure Concentration (ppm)**  Boardstock and Bunstock 302 a Appliance Foam 256 b Laminated Boardstock <1 aConcentrations of HFC - 245fa in this 4 - trial boardstock and bunstock report ranged from <1 ppm (the detection limit) to 7 95 ppm , with the majority of measurements recording the detection limit . The highest concentrations were obtained from one manufacturing facility and were considered not to be commercially acceptable . Thus, it is believed that with proper facility management , these concentrations woul d not be reached . The SolsticeTM 1233zd(E) concentration reported here (i.e., 302 ppm) is modeled based on the maximum reported HFC - 245fa concentration that a person could reasonably be expected to be exposed to for the boardstock and bunstock scenario. b The highest concentration cited in this report on appliance foam exposure HFC - 245fa concentrations was 5,549 ppm, however, this reading was from a vent hole

—a point that would not be accessible to workers, and hence not relevant. The SolsticeTM 1233zd(E) c oncentration reported here (i.e., 256 ppm) is modeled based on the maximum reported HFC - 245fa concentration that a person could reasonably be expected to be exposed to for the appliance foam scenario. Based on the toxicity data for the proposed substitute, the modeling indicates that worker exposure concentrations could reach 302 ppm during manufacture of SolsticeTM 1233zd(E) blown foam, which is slightly above the AEL established by the submitter for So lsticeTM 1233zd(E) . The concentrations reported, however, are based on instantaneous readings, and are not time - weighted over an 8 -hour period. Given that 302 ppm was calculated from the maximum exposure concentration recorded that a person could reasonably be expected to be exposed to , and that exposure concentrations over the time duration were significantly lower than maximum recorded value, it is extremely unlikely that concentrations of SolsticeTM 1233zd(E) would reach or exceed the AEL over an 8 -hour ti me -weighted average. Although exposure data w ere not available for the manufacture of integral skin foam and slabstock foam , which are estimated to have the greatest first year losses (see Table 5) , the actual loss at manufacture associated with integral s kin foam and slabstock foam is anticipated to be higher than boardstock and appliance foam, but less than spray foam (discussed in Section 6.2); the majority of losses are anticipated to occur after manufacture , during processing (Jeffs 2012) . For slabstock, this would include cutting of the foam, and for integral skin foams, this would include removing the foam from its molds. However, the resulting occupational exposure during these activities is not anticipated to present a significant concern (J effs 2012). Further, since the modeled instantaneous concentrations for spray foam application are less than the 8 -hour AEL for SolsticeTM 1233zd(E) , it is unlikely that the time -weighted concentrations during manufacture and processing of integral skin foam and slabstock would result in concentrations beyond the acceptable occupational exposure limit. During manufacture and use of SolsticeTM 1233zd(E), engineering controls should be in place, such as vapor - in air detection systems. Adequate ventilation should always be established during any use, handling, or storage of SolsticeTM 1233zd(E) . In addition, an eye wash and safety shower should be near***** Final - August 3, 2012 *****

the manufacturing facility and ready for use. In general, use of OSHA Category C or higher PPE is recommended, such as respiratory protection (a NIOSH - approved positive -pressure supplied - air respirator is recommended where there is insufficient ventilation; self - contained breathing apparatuses should be used for rescue and maintenance work in storage tanks), chemic al splash goggles or face shield, impervious gloves, and chemical -resistant boo ts and clothing (Anonymous Submitter 2011b , OSHA 1994). Contaminated clothing is not to leave the workplace and must be cleaned before reuse (Anonymous Submitter 2011b ) . During handling of liquid SolsticeTM 1233zd(E) blowing agent for manufacture of blown foam, there is also risk of generation of toxic decomposition products such as hydrochloric acid, hydrofluoric acid, and carbonyl halides when liquid SolsticeTM 1233zd(E) is exposed to fires or temperatures greater than 250°C. Containers of SolsticeTM 1233zd(E) should not be allowed to contact open flames, glowing metal surfaces, or electrical heating elements. EPA believes that because proper handling guidelines are foll owed in accordance with good industrial hygiene and manufacturing practices and the MSDS for SolsticeTM 1233zd(E) , as described above, manufacture of SolsticeTM 1233zd(E) foam does not present a significant risk to workers. 

### 6.2. **Occupational Exposure at**  **Spray Foam Application**  In addition to occupational exposure during manufacture of blown foam, there is also potential for occupational exposure to SolsticeTM 1233zd(E) during rigid polyurethane spray foaming operations , which typically occurs on the site o f installation, outside of a manufacturing facility . To ensure SolsticeTM 1233zd(E) does not present a risk during spray foam application, modeling was again performed for the proposed substitute using the HFC - 245fa monitoring data provided by the submitter as a surrogate . Using the HFC - 245fa surrogate data provided, the maximum possible exposure for employees involved in the use of SolsticeTM 1233zd(E) for spray foams was calculated to be 291 ppm, which is less than the AEL of 300 ppm. Although 291 ppm is close to the AEL for SolsticeTM 1233zd(E) , the surrogate data provided is based on instantaneous readings and are not time - weighted over an 8 -hour period. Given that 291 ppm was calculated from the maximum exposure concentration and that exposure concentrations over the time duration were significantly lower than maximum recorded value , concentrations of SolsticeTM 1233zd(E) would not reach or exceed the AEL over an 8 -hour time -weighted average. Further, the submitter has indicated that use of SolsticeTM 1233z d(E) in the spray foam end -use is anticipated for use only by trained personnel or industrial applications. As such, personnel utilizing SolsticeTM 1233zd(E) in this end -use should ensure that blowing agent losses are minimized and that adequate ventilation is in place. EPA believes that because proper handling guidelines are followed in accordance with good industrial hygiene and manufacturing practices and the MSDS for SolsticeTM 1233zd(E) , use of SolsticeTM 1233zd(E) for the spray foam end -use does not pre sent a significant risk to workers. 

### 7. **GENERAL POPULATION EXPOSURE ASSESSMENT** This section screens potential risks to the general population from exposure to releases of SolsticeTM 1233zd(E) , specifically acute exposures when used in spray foam, and chronic ex posures when released from foams. Because the proper safety precautions, as listed in the remainder of this section, are followed, SolsticeTM 1233zd(E) is not expected to cause a significant impact on human health in the general population when used for foam end -uses. **7.1 Acute Exposure to**  **SolsticeTM 1233zd(E) at Spray Foam Application**  There is potential for general population exposure to SolsticeTM 1233zd(E) during rigid polyurethane***** Final - August 3, 2012 *****

spray foaming operations when performed in buildings with occupants. To ensure that these instances do not present a risk to the general population, EPA recommends that the contractors consult the building management prior to commencing foam-blowing activities . The contractor should also ensure that building occupants are made aware that foam -blowing is about to occur and should suggest the occupants leave the premises until foam-blowing is complete. **7.2 Chronic Exposure to**  **SolsticeTM 1233zd(E) during Lifetime Foam**  **- Blowing Agent Releases**  There is the potential for chronic general population exposures to foam-blowing agents during the lifetime of the foam in which they are used, because these agents can slowly leak from the foam over time. As a result, in locations where persons are regularly present for extended periods, such as a home or apartment, the persons could be exposed to very low levels of SolsticeTM 1233zd(E) over time. Based on the foam blowing end -uses proposed for SolsticeTM 1233zd(E) , the maximum annual rate of release of Sol sticeTM 1233zd(E) during the lifetime of the foam (i.e., after manufacture and before disposal) is anticipated to be 2.5%, resulting in very minimal exposure concentrations. In addition, the submitter has also provided modeling of potential emissions based on the proposed substitute’s diffusion coefficient, and estimated an average SolsticeTM 1233zd(E) concentration of 1.6 ppm within the first two days of a home insulated with spray foam, reducing to 0.008 ppm on average during the foam lifetime (10 - 20 years) . Due to the low toxicity of SolsticeTM 1233zd(E) and minimal expected release, EPA believes that such exposures do not pose a significant toxicity risk, and thus, SolsticeTM 1233zd(E) is not expected to cause a significant impact on human health in the gen eral population when used as a foam blowing agent in the proposed end -uses. 

### 8. **VOLATILE ORGANIC COMPOUND ASSESSMENT** SolsticeTM 1233zd(E) has not been exempted as a volatile organic compound (VOC) under the CAA (40 CFR 51. 1 00 (s) ), and as such, emissions of Sols ticeTM 1233zd(E) should be controlled. EPA has received a petition to exempt this compound from the definition of VOC for purposes of regulations in state implementation plans for local air quality because of low photochemical reactivity that would result in insignificant impacts on ground- level ozone; however, SolsticeTM 1233zd(E) continues to be regulated as a VOC for that purpose unless and until EPA issues a final rule exempting it. An assessment was performed to compare the annual VOC emissions from u se of SolsticeTM 1233zd(E) in the foam blowing end -uses in one year to other anthropogenic sources of VOC emissions. Assuming the submitter’s maximum annual production of SolsticeTM 1233zd(E) for al l proposed foam blowing sector end -uses is entirely released, approximately 245 MT of VOCs would be emitted. This is approximately equal to 2 .0 x10 - 3 percent of all anthropogenic VOC emissions in the U.S. 2 While these emissions may be significant, release of total production for all foam blowing end -uses is extremely unlikely to occur. Thus, an additional analy sis w as performed to determine likely annual VOC emissions from use of SolsticeTM 1233zd(E) in the foam blowing sector. Table 

### 5 summarizes leak estimates and market penetrations from the Vintaging Model for the proposed foam blowing end -use s.

2 Based on the 2010 annual total VOC emissions for the U.S. (EPA 2011).***** Final - August 3, 2012 ***** **Table 5**  **. Foam Release**  **Rates and Market Penetrations**  **by End -**  **Use Foam End**  **- Use**  **Operating Leak Rate**  **First Year Loss**  **a Disposal**  **Loss Lifetime (yrs)**  **Annual**

**Release Rate b**  **Market Share**  Integral Skin Foam 2.50% 95% 0% 2 50% 0.2% PU and PIR Rigid – Boardstock 1% 6% 44% 50 2% 40.6% PU Rigid – Commercial Refrigeration 0.25% 6% 90.25% 15 7% 1.3% PU Rigid – Appliance 0.25% 3.75% 39.88% 14 3% 9.4% PU Rigid – Slabstock 0.75% 37.50% 51.25% 15 7% 0.9% PU Rigid – Spray 1.50% 15% 1% 56 2% 11.6% Source: Vintaging Model (v4.4_3.23.11). **a** First year losses include losses at manufacture and other production steps that occur prior to end - use. **b** Calculated as Leak Rate + ( First Year Loss + Disposal Loss)/Lifetime Assuming that use of SolsticeTM 1233zd(E) is distributed according to existing market share for each proposed foam end -use , and the annual release rates for each of those end-uses, 0.3 MT of VOCs would be expected to be released into the atmosphere. This amount is approximately 2.3x10 - 6 percent of all anthropogenic VOC emissions. In addition, the end -uses with the greatest annual release rate are reasonably expected to occur indoors in a manufacturing or disposal facility, which woul d have engineering control s in place ( e.g., exhaust ventilation , vapor - in - air detection systems , and carbon adsorption systems ) such that actual quantities of SolsticeTM 1233zd(E) emissions would be less than the quantity calculated in this assessment . Further, EPA notes that C3 - C6 hydrocarbons such as cyclopentane, other alternatives sometimes used in the same end uses, are also VOCs. Thus, use of SolsticeTM 1233zd(E) would not necessarily result in greater impacts on local air quality than other available options. As the calculated emissions from the s e analyses are several orders of magnitude smaller than other anthropogenic sources of VOC emissions, EPA believes that through the use of standard industry practices , the environmental impacts of these VO Cs are not considered a significant concern .***** Final - August 3, 2012 *****



### 9. **REFERENCES** Anonymous Submitter 2011a. Response to Incomplete SNAP Submission for SolsticeTM 1233zd(E) . October 20, 2011. Anonymous Submitter 2011b. SNAP Submission to EPA for SolsticeTM 1233zd(E) . May 13, 2011. Anonymous Submitter 2012a. SolsticeTM 1233zd(E) Material Safety Data Sheet. February 28, 2012. Anonymous Submitter . 2012b. Response to A Few More Questions on Foam Uses for SolsticeTM 1233zd(E) . February 6, 2012. Anonymous Submitter . 2012c. Response to Incomplete SNAP Submission for SolsticeTM 1233zd(E) . January 31, 2012. EPA 1994. Significant New Alternatives Policy Technical Background Document: Risk Screen on the Use of Substitutes for Class I Ozone - depleting Substances: Refrigeration and Air Conditioning. Stratospheric Protection Division. March, 1994. EPA 2011. National Emissions Inventory Air Pollutant Emissions Trends Data. Accessed 23 December 2011. Updated October 2011. Available at: <http://www.epa.gov/ttn/chief/trends/index.html #tables > Forster, P., V. Ramaswamy, P. Artaxo, T. Berntsen, R. Betts, D.W. Fahey, J. Haywood, J. Lean, D.C. Lowe, G. Myhre, J. Nganga, R. Prinn, G. Raga, M. Schulz and R. Van Dorland. 2007. Changes in Atmospheric Constituents and in Radiative Forcing. In: Climate Change 2007:The Physical Science Basis. Contribution of Working Group I to the Fourth Assessment Report of the Intergovernmental Panel on Climate Change [Solomon, S., D. Qin, M. Manning, Z. Chen, M. Marquis, K.B. Averyt, M.Tignor and H.L. Miller ( eds.)]. Cambridge University Press, Cambridge, United Kingdom and New York, NY, USA. ICF International. 2012. Determination of an Acceptable Exposure Limit (AEL) for SolsticeTM 1233zd(E) . Prepared for U.S. Environmental Protection Agency. January 23, 2012. Jeffs, Mike. Personal communication between ICF International and Mike Jeffs. April 24 - 26, 2012. OSHA. 1994. General description and discussion of the levels of protection and protective gear. 1910.120 App B. Last updated August 22, 1994. Available at: http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=STANDARDS&p_id=9767 Wang D., Olsen S., Wuebbles D. Undated. “Three - Dimensional Model Evaluation of the Global Warming Potentials for tCFP.” Department of Atmospheric Sciences. University of Il linois, Urbana, IL. Draft Report. Wang D., Olsen S., Wuebbles D. 2011. “Preliminary Report: Analyses of tCFP’s Potential Impact on Atmospheric Ozone.” Department of Atmospheric Sciences. University of Illinois, Urbana, IL. September 26, 2011. WMO (Worl d Meteorological Organization), 2011. Scientific Assessment of Ozone Depletion: 2010 , Global Ozone Research and Monitoring Project

—Report No. 52, 516 pp., Geneva, Switzerland, 2011.***** Final - August 3, 2012 ***** 

## Appendix A: **Determination of an Acceptable Exposure Limit (AEL) for**  **T rans**  **- 1**  **- c**  **hloro -** **3,3,3 -**  **trifluoropro -**  **1 -**  **ene** Recommended AEL: 1,000 ppm (8 - hour time - weighted average) Basis and Endpoints: NOAEC: 4,000 ppm (no significant cardiac histopathology

observed at this concentration) Critical Study : 90 - day inhalation toxicity

study in the rat (Muijser H., 2011) Protocol: Whole -body inhalation, 6 hours/day , 5 days/week for 13 weeks Concentrations: 0; 4,000; 10,000; 15,000 ppm NOAEC 3 

### : 4,000 ppm (identified by independent pathologist’s report [Engelhardt, 2011]) NOAEC [HEC 4 

### ]: 4, 000 ppm x 6 hr/8 hr = 3,000 ppm Uncertainty Factors: 3

– animal to human extrapolation **I. Introduction**  T rans - 1 - chloro - 3,3,3 - trifluoropro - 1 - ene has been proposed as a replacement for PFCs, HCFCs, HFCs and other halogenated compounds. It is intended for several end - uses, including as a foam blowing agent, a refrigerant in chillers, a solvent for cleaning in equipment and aerosol cans, and a carrier solvent in adhesives, coatings, and inks. Several studies assessing the potential toxicity of trans - 1 - chloro - 3,3,3 - trifluoropro - 1 - ene have been conducted and include the following: 14 - day , 28 - day , and 90 - day

inhalation study

in rats, developmental inhalation studies in rats and rabbits, a cardiac sensitization assay in beagle dogs, and genotoxicity studies in bacteria and mammalian cells in culture. The results of these studies are discussed in the following sections to inform an asse ssment of the potential health risks from human exposure to the study compound in the intended end use. **II. Summary of Toxicity Studies**  The toxicological studies reviewed for the determination of acceptable exposure limit for trans 1 - chloro - 3,3,3 - trifluoropro - 1 - ene are summarized below in Table 1. Subclinical effects in the heart (monocy

te infiltration) associated with inhalation exposure to trans - 1 - chloro - 3,3,3 - 3 No Observable Adverse Effect Concentration (NOAEC) 4 Human Equivalent Concentration (HEC)***** Final - August 3, 2012 *****



### trifluoropro - 1 - ene were considered to be the most sensitive human health effects for derivation of an AEL; therefore inhalation studies are described and discussed in detail. The remaining studies are considered supporting studies. **Table 1. T** 

**rans -**  **1 -**  **c hloro**  **- 3,3,3**  **- trifluoropro**  **- 1**  **- ene**  **Toxicological Studies Inhalation Studies**  **Doses NOAEL/**  **NOAEC a**  **Effects** 4 - hour acute (nose - only, rat) 96,000, 156,000 and 120,000 ppm LC 50 , 120,000 ppm (combined sexes) Mortality at  96,000 ppm for some animals. Tremors at all exposures. 14 - day (whole body, rat) 0, 2,000, 7,500, and 20,000 ppm 2,000 ppm (study rpt.)

7,500 (peer review) b Min. focal mononuclear cell infiltrate (heart) 28 - day with 14 - day recovery (nose - only, rat) 0, 2,000, 4,500, 7,500, and 10,000 ppm 4,500 ppm c (study report)

10,000 ppm (peer review) Increased serum K+ in 7,500 - ppm males; reversible changes in hematology/clinical chemistry parameters 90 - day study (nose only, rat) 0, 4,000, 10,000 and 15,000 ppm <4,000 (study rpt.)

4,000 ppm (peer review) Min. focal mononuclear cell infiltrate (heart) Rat fetotoxicity 0, 4,000, 10,000, an d 15,000 ppm 10,000 ppm Dilated urinary bladders in the fetuses Rabbit fetotoxicity 0, 2,500, 10,000, and 15,000 ppm 15,000 ppm No adverse effects noted Cardiac sensitization, dog 25,000, 35,000, and 50,000 ppm Negative at 25,000 ppm Tremors present during exposure  35,000 ppm **Supporting Studies Doses**  **NOAEL/ NOAEC a**  **Effects** In vitro bacterial reverse mutation assay Max concentration of 905,000 ppm NA Negative In vivo unscheduled DNA synthesis 0, 7,500, and 10,000 ppm NA Negative In vitro chromosomal aberration assay 469, 783, or 1,305 μg/mL NA Negative In vivo micronucleus test (rats) 0, 2,000, 4,500, 7,500, and 10,000 ppm NA Negative In vivo micronucleus test (mice) 0, 50,000 ppm NA Negative aAll values NOAEL/NOAEC unless otherwise reported; NA- not applicable b NOAEC values were revised in a pathology peer review report (Engelhardt, 2011) c NOAEC identified in study report based on clinical chemistry, not histopathology results NOAEL = No observed adverse effect level (commonly used for oral or dermal exposures) NOAEC = No observed adverse effect concentration (commonly used for inhalation exposures) **III. Development of the AEL for**  **T rans**  **- 1**  **- c**  **hloro -**  **3,3,3 -** **trifluoropro -**  **1 -**  **ene** Several repeated dose inhalation studies assessing the toxic potential of trans - 1 - c hloro - 3,3,3 -trifluoropro 1 - ene were available for development of an AEL. The NOAEC of 4,000 ppm from the 90 - day inhalation study was chosen as the point of departure for derivation of an AEL for trans - 1 - c hloro - 3,3,3 -trifluoropro 1 - ene as this was the study with the longest exposure duration with an identifiable NOAEC. The AEL was calculated in the following manner: 4000 ppm x (6/8) ÷ 3 = **1000 ppm******* Final - August 3, 2012 *****

A human equivalent concentration (HEC) was determined by adjusting the NOAEC of 4,000 ppm by the ratio of rat exposure duration per day to that of an occupational worker (6 hours/8 hours). The guidelines for developing Reference Concentration (RfC) values (U.S. EPA, 1994) were foll owed, which obviated the need to apply a full uncertainty factor (UF) of 3 for pharmacokinetic differences between rats as the animal model of study and humans (a UF of 3 for pharmacodynamic [PD] differences between the two species has been applied). Becau se subchronic studies are typically used to develop AEL values, an additional UF was not added to account for study duration. Similarly, the database is considered comprehensive, and no additional UF was added to account for database limitations. **IV. Inhala** **tion Studies for T**  **rans -**  **1 -**  **c hloro**  **- 3,3,3**  **- trifluoropro**  **- 1**  **- ene**  **Four -**  **Hour Acute Inhalation Study** In an acute 4 -hour nose - only inhalation study, male and female Crl:SD (outbred) rats (5/sex) were exposed to three different concentrations of trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene (purity 99.99%) and observed for 14 days post - exposure (when possible) (van Triel, 2009). In the first group (Group A), animals were exposed to 100,000 ppm trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene . Because of the low toxicity observed, a second group (Group B) of 5/sex were exposed to a target concentration of 125,000 ppm, that actually reached 156,000 ppm due to technical difficulties; all animals died during exposure. A third group (Group C) of 5 animals/sex were then expo sed to a target concentration of 120,000 ppm in an effort to provide more data to identify the LC 50 . One female from Group A died on the first day of the recovery period. Other animals, male and female, showed clinical signs during recovery, but did not die prematurely. Necropsy did reveal gray discolored lungs in these animals, which was coincident with petechiae in some cases. Three males and two females of Group C died during exposure. Necropsy of these animals revealed reddish lung, with some als o being enlarged. In two female animals of group B, red spots were also found on the thymus. In animals sacrificed on schedule following recovery, no other abnormalities were found. The combined- sex LC 50 was reported as 120,000 ppm. **Fourteen -** 

**Day Inhalation Study**  In a dose -range - finding inhalation study, Crl:CD(SD) rats (5/sex/group) were exposed via whole body inhalation to 0, 2,000, 7,500, or 20,000 ppm trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene (99.9% pure) (Staal, 2008). Exposures were for 6 hr/day, 5 days/week, in a 2 -week period (10 total exposure days). Clinical signs, body weights and food consumption data were collected. Blood samples were drawn at the end of each week for hematology and clinical chemistry analyses. Gross necropsies were p erformed on all animals following the last exposure, and organs were collected and weighed; select organs, including the respiratory tract, were processed and analyzed histologically. The only clinical sign reported during exposure was restlessness, par ticularly in the highest concentration; the effect resolved after 15 minutes of exposure. Test - compound exposure did not induce changes in body weights or food consumption. Regarding changes in hematology, the study author noted an increase in prothrombin time in females at 20,000 ppm, and an increase in absolute and relative numbers of monocytes and absolute number of neutrophils in males at 20,000 ppm. Increases in serum liver enzymes (ALAT 5 and ASAT in males at 20,000 ppm) were not confirmed by any changes in liver histopathology, and were thus considered not toxicologically relevant. Histology revealed multifocal mononuclear cell infiltrates, identified by the study laboratory in animals at 7,500 and 20,000 ppm. However, following an independent pathological review of the histology slides of the heart tissues (Engelhardt, 2011), many of 5 ALAT, alanine transaminase; ASAT, aspartate transaminase***** Final - August 3, 2012 *****

the lesions originally identified by the study laboratory as ‘very slight’ or ‘slight focal myocardial mononuclear cell infiltrate’ were downgraded to being ‘not remarkable’ or were noted to be located only in the apex of the heart. According to the reviewing pathologist, lesions in the apex and/or adjacent portions of the right and left ventricles and interventricular septum of the heart in rats with such lesions are “typical of rodent spontaneous cardiomyopathy”. Given the revised heart data, the reviewing pathologist increased the NOAEC to 7,500 ppm from the original NOAEC of 2,000 ppm identified by the study laboratory. ICF accepts the revised value as reasonable , given the expertise of the reviewing pathologist. **Four -**  **Week Inhalation Study** In a longer -term, nose - only, multi - exposure study, five groups of Sprague - Dawley rats were exposed 6 hrs/day, 5 days/week in a 28 - day period (e.g., 20 - 21 exposure days) to the following nominal concentrations of trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene (99.9% pure): 0, 2,000, 4,500, 7,500, and 10,000 ppm (Staal, 2009). Additional groups of 5/sex were exposed to either 0 or 10,000 ppm in an identical fashion, followed by a 14 - day recovery period. The following parameters were measured in accordance with harmonized guidelines for this repeat - exposure study: clinical signs and mortality, food consumption and body weights, hematology and clinical chemistry, gross necropsy, organ weights, and histopathology of select organs. The study compound did not induce any changes in clinical signs or mortality, food consumption, body weights, urinalysis parameters, gross necropsy, or organ weights. In contrast to the histopathological findings from the 14 - day study, no mononuclear cell infiltration was noted in cardiac tissue or any other selected organ. An increase in the relative number of basophils was observed in males at 10,000 ppm, which resolved in the 14 - day recovery period. Changes in clinical chemistry parameters were limited to decreased creatinine in males at 10,000 ppm, which also was resolved at the end of the recovery period; and an increase in serum potassium in males at both 7,500 and 10,000 ppm, which were still slightly ele vated after recovery, although not statistically significant. The study authors identified a NOAEC of 4,500 ppm, arguing that the increased potassium levels at exposure concentrations of 7,500 and 10,000 ppm were related to treatment. The pathologist who performed the peer review of the cardiac histopathology slides for this study identified a NOAEC of 10,000 ppm, based on the confirmed absence of compound - induced subcellular damage to cardiac tissue. ICF concurs with the reviewing pathologist, and would add that although the increased potassium level observed at ≥7,500 ppm may be indicative of subclinical cardiotoxicity, it cannot be attributed to the test compound alone, and may also be attributed to spontaneous cardiomyopathy. The NOAEC for this study is set, therefore, at 10,000 ppm. **Ninety -**  **Day Inhalation Study** In a 90 - day inhalation study, male and female Sprague - Dawley (Crl:CD[SD]) rats (10/sex/concentration) were exposed to trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene (99.9% purity) via nose - only exposu re at nominal vapor concentrations of 0, 4,000, 10,000, or 15,000 ppm for 6 hrs/day for 5 days/week for 13 weeks, for a total of 65 exposure days (Muijser, 2011). Rats were observed twice daily for clinical signs of toxicity, morbidity, and mortality. Bo dy weights and mean food consumption were recorded weekly. Ophthalmoscopic examinations were conducted at study initiation and during the last week of exposure. Blood samples and urine samples were taken just prior to or the last day prior to sacrifice at the end of the exposure period and were analyzed for hematology, clinical chemistry and urinalysis parameters consistent with harmonized guidelines. Gross necropsies were performed on all surviving animals after the scheduled exposure period, and select o rgans were removed, weighed, and prepared for histopathology.***** Final - August 3, 2012 *****

Mortality, Clinical Effects, Eye Effects, Food Consumption, Body Weights Exposure to the study compound did not induce early mortality or any observed clinical abnormalities. In addition, exposure did not affect body weight gain, food consumption, or changes in eye condition. Hematology/Clinical Chemistry Effects on hematological parameters were limited. For example, exposure to the study compound had no effect on red blood cell parameter s and affected white blood cells only by decreasing the relative number of lymphocytes in males at 15,000 ppm. Clinical chemistry effects were inconsistent between the sexes. Plasma of ASAT and ALAT were increased at statistically significant levels in males at 15,000 ppm, but not in females, while glucose and urea were increased in females at 10,000 and 15,000 ppm, but not in males. High - concentration females also exhibited significantly increased potassium and decreased triglyceride levels. No other c hanges in measured analytes were noted. Organ Weights A treatment -related decrease in absolute and relative heart weights was noted in male rats, which reached statistical significance at the highest concentration (15,000 ppm). In addition, relative liver weights in the males were significantly increased at the highest concentration, whereas by contrast, the relative liver weights of high - concentration females were significantly decreased. No other changes in organ weights were noted. Macroscopic and Microscopic Findings Exposure to trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene did not induce any changes in the appearance of organs or tissues in the bodies of rats evaluated following gross necropsy. In general, no compoundinduced effects were noted upon analyses of histopathology findings, with the exception of minimal multifocal mononuclear cell infiltrate in the heart. The study authors also identified inflammation occurring coincident with the infiltrates in some animals. The report of the peer review pathologist indicated that the vast majority of these findings were downgraded to categories of ‘not remarkable’ or were localized to the apex, the lower left ventricle, or the atrium. Again, because of the downgrading of incidences of infiltration reported in the original study, and the identification of some mononuclear infiltration as ‘spontaneous cardiomyopathy,’ the NOAEC was identified as the lowest exposure concentration of 4,000 ppm (the study authors reported the NOAEC as less than 4,000 ppm). ICF concurs, and as such, the NOAEC used in the determination of an AEL for trans - 1 - c hloro - 3,3,3 - trifluoropro - 1 - ene is 4,000 ppm. **Developmental Toxicity** T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was evaluated in two developmental toxicity studies, one in rats and one in rabbits. Rat Prenatal Developmental Study In the prenatal developmental study in rats (Waalkens - Berendsen and van den Hoven, 2011), groups of 24 mated female Wistar (Crl:WI[WU] ) rats were exposed 6 hrs/day to concentrations of 0, 4,000, 10,000 or 15,000 ppm trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene (99.9% pure) from gd 6 to gd 19. Health parameters measured during the in - life phase of the study included mortality, clinical signs, maternal body weight, and food consumption. A Caesarean section was performed following the gestation period, and females and fetuses were evaluated via gross necropsy, and for standard reproductive toxicity endpoints (e.g., gestation index, fecundity index, fetus sex ratio, number of corpora lutea, implantation sites, pre - and post - implantation loss, live and dead fetuses, and resorptions). Fetuses, placentas and reproductive organs were weighed, and fetuses were processed for soft tissue and skeletal a bnormalities and malformations.***** Final - August 3, 2012 *****

Exposure to the study compound did not affect any of the measured parameters with the exception of the following: a statistically significant increase in the incidence of dilated urinary bladders was observed in the fetuses of the highest concentration group (15,000 ppm). Because the incidence of this anomaly increased with higher concentrations, it is considered treatment -related. No signs of maternal toxicity were noted at any concentration. The NOAEC for developmental effects is established at 10,000 ppm; the NOAEC for maternal toxicity is established at 15,000 ppm. Rabbit Prenatal Developmental Study T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was further evaluated in a developmental study in the New Zealand white rabbit. Hoffman (2010) exposed 22 pregnant dams/group during organogenesis (gd 6 to gd 28) to trans - 1 - c hloro - 3,3,3 - trifluoropro - 1 - ene (~100% purity) via whole -body inhalation for 6 h rs/day at concentrations of 0, 2,500, 10,000 or 15,000 ppm. During the in - life phase of the study, the rabbits were observed for mortality, clinical signs, body weights, and food consumption. Following euthanasia of the dams at gd 29, the following parameters were measured: gross necropsy, number of corpora lutea and implantation sites, uterine and placental weights, fetus number (live), fetal weights (live only), sex ratio, external defects, soft tissue abnormalities, and skeletal abnormalities and state of ossification. Exposure to the test compound did not affect any changes in any of the measured parameters of the exposed dams or the fetuses. Therefore, the NOAEC for both maternal and fetal toxicity is established at 15,000 ppm in this rabbit devel opmental assay. **Cardiac Sensitization** In a muzzle - only, acute exposure study in young beagle dogs (Atterson, 2011), 6 male dogs were exposed successively to 25,000 ppm (2.5%), 35,000 ppm (3.5%) and 50,000 ppm (5%) trans - 1 - c hloro - 3,3,3 - trifluoropro - 1 - ene ( 99.978% pure) up to a maximum period of 33 minutes, with 48 hours of rest occurring in between exposure periods. Control (air) exposures occurred in the same dogs. For the cardiac sensitization, epinephrine challenge (bolus) injections, ranging from 2 to 8 μg/kg, were started approximately 5 minutes following the initiation of exposure and continued until termination of exposure. Only 2 of the 6 dogs were exposed to the highest trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene concentration, because behavioral resp onses (vocalization, injected sclera, excessive salivation, tremors, convulsions and/or reddened gums) prompted the termination of exposure; only one of the dogs at this concentration was given an epinephrine challenge. No test -related deaths occurred in the study. Clinical findings similar to those at 50,000 ppm were noted after exposure to 35,000 ppm (tremors, injected sclera, reddened ears and excessive salivation). No signs of cardiac sensitization were noted at 25,000 or 35,000 ppm; however, the pres ence of ECG noise in two animals exposed to 35,000 ppm precluded the complete characterization of cardiac sensitization at this concentration. As noted previously, exposure was terminated at the highest concentration and assessment of cardiac sensitizatio n was not possible at this level. Therefore, the results indicate that the study compound does not cause cardiac sensitization at 25,000 ppm.***** Final - August 3, 2012 *****

**V. Supporting Genotoxicity Studies for**  **T rans**  **- 1**  **- c**  **hloro -**  **3,3,3 -**  **trifluoropro -**  **1 -**  **ene Reverse Mutation Assay**  T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was tested in an Ames bacterial reversion assay in Salmonella typhimurium (S. typhimurium ) strains TA98, TA100, TA1535, TA1537, and E. coli strain WP2 uvrA (Wagner et al., 2011). This assay measures the ability of the compound to induce either base pair or frame shift mutations in DNA in the presence or absence of S9 fraction from induced rat livers (metabolizing enzymes including mixed function oxidases). The study compound was analyzed for mutagenicity twice at concentrati ons ranging from 73,400 to 905,000 ppm at 25°C for either 24 or 48 hours. Appropriate negative and positive controls were analyzed in the same assays. Mutagenicity was not induced in any strain, but toxicity was seen in all strains at the top two concent rations of 513,000 and 905,000 ppm. A confirmatory assay was performed using slightly reduced exposure concentrations of 17,200; 24,500; 73,400; 147,000; 196,000; 269,000; 513,000 and 905,000 ppm at 25°C. Cytotoxicity was observed at the top two concentrations, as in the previous assays. No positive responses were observed with strains TA98 and TA1537 in the absence of S9 activation and with strains TA98 and WP2uvrA in the presence of S9 activation. The compound is categorized as negative in this genoto xicity assay. **Unscheduled DNA Synthesis** T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was tested for the ability to induce unscheduled DNA synthesis in liver hepatocytes isolated from male rats exposed to 7,500 and 10,000 ppm of the study compound in a 28 - day study (Staal, 2009). The control group was the same one used in the 28 - day study and positive control animals were dosed with standard toxicants per harmonized guidelines. T rans - 1 - c hloro - 3,3,3 - trifluoropro - 1 - ene did not induce unscheduled DNA synthesis under the conditions used in the 28 - day study, and is categorized as negative for this genotoxicity assay. **Micronuclei** Two studies were performed to assess the ability of trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene to include micronuclei in bone marrow, as described below. Micronuclei Rat Study T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was tested for the ability to induce micronuclei in bone marrow cells isolated from male rats exposed to 0, 2,000, 4,500, 7,500 and 10,000 ppm of the study compound in a 28 - day study (Staal, 2009). Appropriate positive control animals were dosed with sta ndard toxicants per harmonized guidelines. The test compound did not induce chromosomal damage or induce micronuclei in the bone marrow under the conditions used in this study (the maximum concentration administered, 10,000 ppm is equivalent to >12,000 mg/kg, much higher than the 1,000 mg/kg limit dose recommended by EPA) and is judged negative for this genotoxicity assay. Micronuclei Mice Study T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was tested in an independent assay for its ability to induce micronucle i in the bone marrow of male mice (de Vogel, 2009). Two groups of 10 male, albino mice (CD - 1 strain, Charles River) were exposed for a single, nose - only, 4 -hour exposure to trans - 1 - c hloro 3,3,3 -trifluoropro - 1 - ene (99.99% pure) at concentrations of 0, or 5 0,000 ppm. Five male mice were injected intraperitoneally with 0.75 mg/kg -bw of mitomycin C as a positive control for comparison. Bone marrow cells were isolated from half the exposure group at 24 hours and the remainder at 48 hours postexposure; bone marrow smears were prepared and analyzed for the presence of polychromatic and normochromatic erythrocytes and micronuclei in a manner consistent with established guidelines. The test compound did not induce micronuclei or any other form of chromosomal damage and was thus judged negative in this genotoxicity assay.***** Final - August 3, 2012 *****

**Chromosomal Aberration** T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene (99.98% pure) was tested in a chromosomal aberration assay with human lymphocytes cultured in vitro (Pritchard and Damant, 2011). Human lymphocytes, isolated from whole blood samples provided by volunteers, were treated with trans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene for either 3 or 21 hours at doses of 469, 783, or 1,305 μg/mL in the presence or absence of metabolic activation (S9 from the livers of chemically- induced rats). T rans - 1 - c hloro - 3,3,3 -trifluoropro - 1 - ene was determined to be negative in this genotoxicity assay, as the cells treated with trans - 1 - c hloro - 3,3,3 - trifluoropro - 1 - ene in all experiments did not cause an increase in the frequency of cells exhibiting structural chromosomal aberrations compared with negative controls at any concentration tested. **References** Atterson PR. 2011. Acute cardiac sensitization study of 1233zd in beagle dogs. WIL Research Laboratories, LLC, Ashland, OH, performed for Anonymous Submitter , NJ . 21 April 2011. de Vogel N and van Triel JJ. 2009. Micronucleus test in bone marrow cells of mice treated with 1233zd(E), administered by inhalation. TNO Quality of Life , The Netherlands, performed for Anonymous Submitter , Morristown, NJ, 10 February 2009. Engelhardt JA. 2011. Peer review of hearts from inhalation toxicity studies with various fluorocarbons. Experimental Pathology Laboratories, Inc. 19 October 2011. Hof fman G. 2010. 1233zd(E): Embryo - fetal toxicity study in rabbits via whole -body inhalation exposure (GLP). Huntingdon Life Sciences, New Jersey, for Anonymous S u bmitter New Jersey. 2 September 2010. Muijser H. 2011. Sub - chronic (13 -week) inhalation toxic ity study with 1233zd(E) in rats. TNO Quality of Life, The Netherlands, performed for Anonymous Submitter , NJ , 3 May 2011. Pritchard L and Damant C. 2011. 1233zd in vitro mammalian chromosome aberration test in human lymphocytes. Amended Final Report. Huntingdon Life Sciences UK for Anonymous Submitter . NJ, 31 March 2011. Staal YCM. 2009. A sub - acute (4 -week) inhalation toxicity study, including unscheduled DNA synthesis and micronucleus test, with 1233zd(E) in rats. TNO Quality of Life, The Netherlands, performed for Anonymous Submitter , USA, 27 May 2009. Staal YCM. 2008. Sub - acute (14 - day) inhalation toxicity study with 1233zd(E) in rats. TNO Quality of Life, The Netherlands, performed for Anonymous S ubmitter , USA, 15 December 2008. U.S. EPA. 1994. Methods for Derivation of Inhalation Reference Concentrations (RfCs) and Application of Inhalation Dosimetry. U.S. Environmental Protection Agency, Office of Research and Development, Office of Health and Environmental Assessment, Washington, DC, EPA/600/8 - 90/066F. van Triel JJ. 2009. Acute (4 -hour) inhalation toxicity study with 1233zd(E) in rats. TNO Quality of Life, The Netherlands, performed for Anonymous Submitter , USA, 17 February 2009. Waalkens - Berendsen DH, and MJW van den Hoven. 2011. A prenatal developmental inhalation toxicity study with 1233zd(E) in rats. TNO Triskelion, The Netherlands, for Anonymous Submitter , New Jersey, 28 January 2011. Wagner VO, Hines RM, Jois M. 2008. 1233zd(E) bacterial reverse mutation assay using gas -phase e xposure. Amended Final Report. BioReliance, Rockville, MD, performed for Anonymous Submitter , New Jersey . 27 April 2011.